4-epi-Isofagomine derivatives as pharmacological chaperones for the treatment of lysosomal diseases linked to β-galactosidase mutations: Improved synthesis and biological investigations.
(5aR)-5a-C-pentyl-4-epi-isofagomine 1 is a powerful inhibitor of lysosomal β-galactosidase and a remarkable chaperone for mutations associated with GM1-gangliosidosis and Morquio disease type B. We report herein an improved synthesis of this compound and analogs (5a-C-methyl, pentyl, nonyl and phenylethyl derivatives), and a crystal structure of a synthetic intermediate that confirms its configuration resulting from the addition of a Grignard reagent. These compounds were evaluated as glycosidase inhibitors and their potential as chaperones for mutant lysosomal galactosidases determined. Based on these results and on docking studies, the 5-C-pentyl derivative 1 was selected as the optimal structure for further investigations: this compound induces the maturation of mutated β-galactosidase in fibroblasts of a GM1-gangliosidosis patient and promote the decrease of keratan sulfate and oligosaccharide load in patient cells. Compound 1 is clearly capable of restoring β-galactosidase activity and of promoting maturation of the protein, which should result in significant clinical benefit. These properties strongly support the development of compound 1 for the treatment of GM1-gangliosidosis and Morquio disease type B patients harboring β-galactosidase mutations sensitive to pharmacological chaperoning.


WE-G-211-02: Origins and Development of Magnetic Resonance Imaging.
The method of magnetic resonance imaging (MRI) was an extension of nuclear magnetic resonance (NMR) spectroscopy that was enabled by computational systems capable of performing the Fast Fourier Transform. Initially, a diverse group of methods were proposed for producing images with NMR, with the multi-slice spin-warp spin echo MRI method winning out as the first clinically useful approach. Immediately radiologists were impressed in the soft tissue contrast seen in MR images and the great flexibility with which image contrast could be manipulated. Over the years, advances in both hardware and the implementation of new pulse sequences of increasing complexity led to the increasing utility of MRI. The great success of MRI research contributed to the awarding of both the Nobel Prize in Chemistry for Richard Ernst in 1991 and the Nobel Prize in Physiology or Medicine for Lauterbur and Mansfield in 2003. The evolution of the modality produced increasing speed of acquisition, finer spatial resolution and an expanding range of clinical applications. Technological advances included improvements in gradient coils, superconducting magnets, phased array radiofrequency coils, and partially parallel image reconstruction algorithms. The wide range of applications developed include MR angiography, BOLD contrast functional MRI, and diffusion-weighted MRI. This section of the symposium will conclude with a look a current developments and where they are likely to take the field into the future.


Computational imaging of the cardiac activities using magnetocardiography.
The electrical impulses of the heart will generate a tiny magnetic field outside the thorax that is measured as Magnetocardiographic signals. The challenging study is to estimate the cardiac activities in terms of depolarisation and repolarization maps from the measured signals called as inverse problem. This is computed only if one has solved generic or subject- specific prior models using the anatomical structures of the myocardium, the torso and the detectors called as forward problem. In this study, the Discretised heart is priorily assumed as the dipolar sources forming a double layer. The thorax structure modelled with finite element meshes is considered in the forward study. The magnetocardiographic data are simulated using uniform double layer model representing transmembrane distribution on the epicardium and endocardium. Using this data, the activation maps are non-invasively imaged on the heart surface using Tikhonov's regularisation technique. The inverse study is extended to reconstruct the depolarisation sequences of the abnormal cases.


Adolescents' perceptions of print cigarette advertising: a case for counteradvertising.
Cigarette smoking is a major preventable cause of serious chronic disease. The majority of smokers begin smoking during their adolescent years. Print cigarette advertising plays an influential role in encouraging young people to smoke. Eight hundred and forty-three middle/junior high-school-aged students from a large midwestern metropolitan area participated in this study of adolescents' recognition of and belief in messages associated with print cigarette advertising. Results indicated no significant difference between smokers and nonsmokers in their recognition of messages associated with print cigarette advertising. However, smokers and nonsmokers differed significantly in the degree to which they believed the messages communicated by print cigarette advertising. Consistent with social cognitive theory, students who smoke at least occasionally were more likely to believe messages conveyed by print cigarette ads than were students who had never smoked. Implications of these findings for research and practice are discussed, including specification of guidelines for effective counteradvertising.


Dynamic vs. static-stretching warm up: the effect on power and agility performance.
The purpose of this study was to compare the effect of a dynamic warm up (DWU) with a static-stretching warm up (SWU) on selected measures of power and agility. Thirty cadets at the United States Military Academy completed the study (14 women and 16 men, ages 18-24 years). On 3 consecutive days, subjects performed 1 of the 2 warm up routines (DWU or SWU) or performed no warm up (NWU). The 3 warm up protocols lasted 10 minutes each and were counterbalanced to avoid carryover effects. After 1-2 minutes of recovery, subjects performed 3 tests of power or agility. The order of the performance tests (T-shuttle run, underhand medicine ball throw for distance, and 5-step jump) also was counterbalanced. Repeated measures analysis of variance revealed better performance scores after the DWU for all 3 performance tests (p < 0.01), relative to the SWU and NWU. There were no significant differences between the SWU and NWU for the medicine ball throw and the T-shuttle run, but the SWU was associated with better scores on the 5-step jump (p < 0.01). Because the results of this study indicate a relative performance enhancement with the DWU, the utility of warm up routines that use static stretching as a stand-alone activity should be reassessed.


Regulation of endodermal differentiation of human embryonic stem cells through integrin-ECM interactions.
Many cellular responses during development are regulated by interactions between integrin receptors and extracellular matrix proteins (ECMPs). Although the majority of recent studies in human embryonic stem cell (hESC) differentiation have focused on the role of growth factors, such as FGF, TGFβ, and WNT, relatively little is known about the role of ECMP-integrin signaling in this process. Moreover, current strategies to direct hESC differentiation into various lineages are inefficient and have yet to produce functionally mature cells in vitro. This suggests that additional factors, such as ECMPs, are required for the efficient differentiation of hESCs. Using a high-throughput multifactorial cellular array technology, we investigated the effect of hundreds of ECMP combinations and concentrations on differentiation of several hPSC lines to definitive endoderm (DE), an early embryonic cell population fated to give rise to internal organs such as the lung, liver, pancreas, stomach, and intestine. From this screen we identified fibronectin (FN) and vitronectin (VTN) as ECMP components that promoted DE differentiation. Analysis of integrin expression revealed that differentiation toward DE led to an increase in FN-binding integrin α5 (ITGA5) and VTN-binding integrin αV (ITGAV). Conditional short hairpin RNA-mediated knockdown of ITGA5 and ITGAV disrupted hESC differentiation toward DE. Finally, fluorescence-based cell sorting for ITGA5 and ITGAV significantly enriched cells with gene expression signatures associated with DE, demonstrating that these cell surface proteins permit isolation and enrichment of DE from hESCs. These data provide evidence that FN and VTN promote endoderm differentiation of hESCs through interaction with ITGA5 and ITGAV, and that ECMP-integrin interactions are required for hESC differentiation into functionally mature cells.


Clinically recognized dysplastic nevi. A central risk factor for cutaneous melanoma.
To investigate the relationship of number and type of nevi to the development of melanoma. Case-control study. Outpatient clinics in referral hospitals. Cases were 716 consecutive patients with newly diagnosed melanoma identified at 2 melanoma centers between January 1, 1991, and December 31, 1992. Stratified random sampling of patients from outpatient clinics was used to identify 1014 participating controls of the same age, sex, race, and geographic distribution as the melanoma cases. All study subjects underwent an interview, a complete skin examination, photography of the most atypical nevi, and, if the patient was willing, a biopsy of the most atypical nevus. Number and type of nevi on the entire body were systematically reported. All diagnoses of clinically dysplastic nevi were confirmed by expert examiners. Risk for melanoma was strongly related to number of small nevi, large nondysplastic nevi, and clinically dysplastic nevi. In the absence of dysplastic nevi, increased numbers of small nevi were associated with an approximately 2-fold risk, and increased numbers of both small and large nondysplastic nevi were associated with a 4-fold risk. One clinically dysplastic nevus was associated with a 2-fold risk (95% confidence interval, 1.4-3.6), while 10 or more conferred a 12-fold increased risk (95% confidence interval, 4.4-31). Congenital nevi were not associated with increased risk of melanoma. Although nondysplastic nevi confer a small risk, clinically dysplastic nevi confer substantial risk for melanoma. On the basis of nevus number and type, clinicians can identify a population at high risk of this epidemic cancer for screening and intervention.


A perspective on stem cell therapy for ear disorders.
The use of stem cells in cell-based therapy is an emerging concept for the treatment of ear disorders. Tympanic membrane perforation (TMP) and inner ear disorders are some of the most commonly presented otologic disorders that can benefit from advances in cell-based therapy. Studies have already demonstrated that stem cell-based therapy can potentially be an effective treatment modality for acute and chronic TMP. Recent studies have also shown promise in application of cell-based approach to treat inner ear dysfunction. In this perspective, we will discuss the recent advancements regarding the use of cell-based therapy for ear disorders.


Developing Capability: Transforming Massage Therapy Education through Inquiry-based Learning.
Massage therapy education programs are responsible for preparing graduates for complex professional environments, while also ensuring graduates can access the profession by passing the regulatory body's standardized certification or licensing examination. Massage therapists are regulated health care professionals in four Canadian provinces. They are autonomous practitioners and an integral part of the health care regime for patients with complicated health presentations. As such, they must think critically about patients' presentations and determine the most appropriate care. This article describes the process undertaken by the faculty team of Humber College's Massage Therapy Program in an effort to transform the existing teaching and learning strategies by incorporating principles of inquiry-based learning (IBL).


Canine Lat1: molecular structure, distribution and its expression in cancer samples.
A full-length cDNA sequence of canine L-type amino acid transporter 1 (Lat1) was determined from a canine brain. The sequence was 1828 bp long and was predicted to encode 485 amino acid polypeptides. The deduced amino acid sequence of canine Lat1 showed 93.2% and 91.1% similarities to those of humans and rats, respectively. Northern blot analysis detected Lat1 expression in the cerebellum at 4 kb, and Western blot analysis showed a single band at 40 kDa. RT-PCR analysis revealed a distinct expression of Lat1 in the pancreas and testis in addition to the cerebrum and cerebellum. Notably, Lat1 expression was observed in the tissues of thyroid cancer, melanoma and hemangiopericytoma. Although the cancer samples examined were not enough, Lat1 may serve as a useful biomarker of cancer cells in veterinary clinic.


Using a therapeutic companion robot for dementia symptoms in long-term care: reflections from a cluster-RCT.
We undertook a cluster-randomised controlled trial exploring the effect of a therapeutic companion robot (PARO) compared to a look-alike plush toy and usual care on dementia symptoms of long-term care residents. Complementing the reported quantitative outcomes , this paper provides critical reflection and commentary on individual participant responses to PARO, observed through video recordings , with a view to informing clinical practice and research. A descriptive, qualitative design with five participants selected from the PARO intervention arm of the trial. The trial is registered with the Australian New Zealand Clinical Trials Registry (ACTRN12614000508673). The five participants and their responses to PARO are presented in terms of three issues: i.) Different pre-intervention clinical presentations and different responses; ii.) Same individual, different response - the need for continual assessment and review; and iii.) The ethics of giving and retrieving PARO. Implications for clinical practice and future research are discussed in relation to each issue. The findings suggest that one approach does not fit all, and that there is considerable variation in responses to PARO. A number of recommendations are discussed to aid the delivery of psychosocial interventions with PARO in practice, as well as to guide future research.


How important is dosimetrist experience for intensity modulated radiation therapy? A comparative analysis of a head and neck case.
Treatment planning for IMRT is a complex process that requires additional training and expertise. The aim of this study was to compare and analyze IMRT plans generated by dosimetrists with varying levels of IMRT planning experience. The computed tomography (CT) data of a patient previously treated with IMRT for left tonsillar carcinoma were used. The patient's preexisting planning target volumes (PTVs) and all organs at risk were provided with the CT data set. Six dosimetrists with variable IMRT planning experience generated IMRT plans according to the department's protocol. Plan analysis included visual inspection and comparison of dose-volume histogram, conformity indices, treatment delivery efficiency, and dose delivery accuracy. Visual review of the dose distribution showed that the 6 plans were comparable. However, only the 2 most experienced dosimetrists were able to meet the strict PTV aims and critical structure constraints. The least experienced dosimetrist had the worst planning outcome. Comparison of delivery efficiency showed that the number of segments, total monitor units, and treatment time increased as the IMRT planning experience decreased. Dosimetrists with higher levels of IMRT planning experience produced a better quality head and neck IMRT plan. Different planning experience may need to be considered when organizing appropriate departmental resources.


Urbanisation but not biomass fuel smoke exposure is associated with asthma prevalence in four resource-limited settings.
Urbanisation is an important contributor to the prevalence of asthma worldwide, and the burden of this effect in low-income and middle-income countries undergoing rapid industrialisation appears to be growing. We sought to characterise adult asthma prevalence across four geographically diverse settings in Peru and identify both individual and environmental risk factors associated with adult asthma. We collected sociodemographics, clinical history and spirometry in adults aged ≥35 years. We defined asthma as meeting one of the three criteria: physician diagnosis, self-report of wheezing attack or use of asthma medications. We used multivariable logistic regression to assess individual and environmental factors associated with adult asthma. We analysed data from 2953 participants (mean age 55 years; 49% male). Overall asthma prevalence was 7.1%, which varied with urbanisation: highest in Lima (14.5%), followed by urban Puno (4.0%), semiurban Tumbes (3.8%) and rural Puno (1.8%). In multivariable analysis, being male (OR=0.60, 95% CI 0.39 to 0.93) and living at high altitude (OR=0.26, 95% CI 0.16 to 0.42) were associated with lower odds of having asthma, whereas living in an urban setting (OR=4.72, 95% CI 3.15 to 7.23) and family history of asthma (OR=1.83, 95% CI 1.19 to 2.73) were associated with higher odds. Current daily exposure to biomass fuel smoke (OR=1.18, 95% CI 0.70 to 1.91) and smoking (OR=0.99, 95% CI 0.73 to 1.22) were not associated with asthma. These findings confirm that urbanisation is an environmental risk factor of asthma, questions biomass fuel smoke exposure as an important risk factor and proposes high altitude as possibly protective against the development of asthma.


Phase I/II study of the hypoxia-activated prodrug PR104 in refractory/relapsed acute myeloid leukemia and acute lymphoblastic leukemia.
We previously demonstrated vast expansion of hypoxic areas in the leukemic microenvironment and provided a rationale for using hypoxia-activated prodrugs. PR104 is a phosphate ester that is rapidly hydrolyzed in vivo to the corresponding alcohol PR-104A and further reduced to the amine and hydroxyl-amine nitrogen mustards that induce DNA cross-linking in hypoxic cells under low oxygen concentrations. In this phase I/II study, patients with relapsed/refractory acute myeloid leukemia (n=40) after 1 or 2 prior treatments or acute lymphoblastic leukemia (n=10) after any number of prior treatments received PR104; dose ranged from 1.1 to 4 g/m(2). The most common treatment-related grade 3/4 adverse events were myelosuppression (anemia 62%, neutropenia 50%, thrombocytopenia 46%), febrile neutropenia (40%), infection (24%), and enterocolitis (14%). Ten of 31 patients with acute myeloid leukemia (32%) and 2 of 10 patients with acute lymphoblastic leukemia (20%) who received 3 g/m(2) or 4 g/m(2) had a response (complete response, n=1; complete response without platelet recovery, n=5; morphological leukemia-free state, n=6). The extent of hypoxia was evaluated by the hypoxia tracer pimonidazole administered prior to a bone marrow biopsy and by immunohistochemical assessments of hypoxia-inducible factor alpha and carbonic anhydrase IX. A high fraction of leukemic cells expressed these markers, and PR104 administration resulted in measurable decrease of the proportions of hypoxic cells. These findings indicate that hypoxia is a prevalent feature of the leukemic microenvironment and that targeting hypoxia with hypoxia-activated prodrugs warrants further evaluation in acute leukemia. The trial is registered at clinicaltrials.gov identifier: 01037556.


Lower hemoglobin transfusion trigger is associated with higher mortality in patients hospitalized with pneumonia.
Patients hospitalized with pneumonia may require packed red blood cell (RBC) transfusion during their hospital stay. Patient survival may be associated with the transfusion trigger. These patients may need a higher hemoglobin (Hb) trigger than that suggested by the AABB guidelines (7 g/dL).The objective of this study was to evaluate the association between the initial transfusion Hb trigger and in-hospital mortality.A historical cohort study of all patients hospitalized in an internal medicine ward between 2009 and 2014 with pneumonia, who received at least 1 unit of RBC, was evaluated. The primary outcome was all-cause in-hospital mortality.One hundred males and 77 females with a median age of 80 (interquartile range 71-87) years were included. The median Hb trigger was 8.10 g/dL. Mortality rate was 56% in patients with Hb trigger ≤7 g/dL, 43.8% in Hb trigger 7 to 8 g/dL, and 29.5% in Hb trigger >8 g/dL (P = .045). Patients in the 3 Hb trigger categories did not differ in age, sex, comorbidities, albumin, creatinine, C-reactive protein, white blood cells, and platelet counts. The result of a multivariate analysis showed that only lower Hb trigger (odds ratio [OR]≤ 7vs.>8 = 5.24, OR7-8vs.>8 = 2.13, P = .035) and higher neutrophil count (P = .012) were associated with increased in-hospital mortality.In conclusion, a lower transfusion trigger is associated with increased risk for in-hospital mortality in patients hospitalized with pneumonia requiring RBC transfusion.


Analgesic and antiinflammatory activities of the essential oil of the unique plant Zhumeria majdae.
The analgesic and anti-inflammatory properties of Zhumeria majdae essential oil were investigated in mice and rats. The analgesic activity of the oil was assessed by acetic acid-induced writhing and Eddy's hot plate methods while the acute anti-inflammatory effect was investigated by inflammatory paw edema in rat. Z majdae oil significantly decreased the number of acetic acid-induced writhes in mice compared with animals that received vehicle only. Also it exhibited a central analgesic effect as evidenced by a significant increase in reaction time in the hot plate method. The oil also significantly reduced carrageenan induced paw edema in rats. The inhibitory activity of Z. majdae essential oil was found to be very close to that of the standard drug, diclofenac sodium (50 mg/kg). The studied oil was analyzed by GC and GC-MS and seventeen constituents were identified, representing 99.2% of the oil. The major components of the oil were characterized as linalool (63.4%) and camphor (27.5%), which might be responsible for these observed activities. The results suggest that Z. majdae essential oil possesses biologically active constituent(s) that have significant activity against acute inflammation and have central and peripheral antinociceptive effects which support the ethnomedicinal claims of the plant application in the management of pain and inflammation.


Multiparametric prostate MR imaging with T2-weighted, diffusion-weighted, and dynamic contrast-enhanced sequences: are all pulse sequences necessary to detect locally recurrent prostate cancer after radiation therapy?
To compare diagnostic accuracy of T2-weighted magnetic resonance (MR) imaging with that of multiparametric (MP) MR imaging combining T2-weighted imaging with diffusion-weighted (DW) MR imaging, dynamic contrast material-enhanced (DCE) MR imaging, or both in the detection of locally recurrent prostate cancer (PCa) after radiation therapy (RT). This retrospective HIPAA-compliant study was approved by the institutional review board; informed consent was waived. Fifty-three men (median age, 70 years) suspected of having post-RT recurrence of PCa underwent MP MR imaging, including DW and DCE sequences, within 6 months after biopsy. Two readers independently evaluated the likelihood of PCa with a five-point scale for T2-weighted imaging alone, T2-weighted imaging with DW imaging, T2-weighted imaging with DCE imaging, and T2-weighted imaging with DW and DCE imaging, with at least a 4-week interval between evaluations. Areas under the receiver operating characteristic curve (AUC) were calculated. Interreader agreement was assessed, and quantitative parameters (apparent diffusion coefficient [ADC], volume transfer constant [K(trans)], and rate constant [k(ep)]) were assessed at sextant- and patient-based levels with generalized estimating equations and the Wilcoxon rank sum test, respectively. At biopsy, recurrence was present in 35 (66%) of 53 patients. In detection of recurrent PCa, T2-weighted imaging with DW imaging yielded higher AUCs (reader 1, 0.79-0.86; reader 2, 0.75-0.81) than T2-weighted imaging alone (reader 1, 0.63-0.67; reader 2, 0.46-0.49 [P ≤ .014 for all]). DCE sequences did not contribute significant incremental value to T2-weighted imaging with DW imaging (reader 1, P > .99; reader 2, P = .35). Interreader agreement was higher for combinations of MP MR imaging than for T2-weighted imaging alone (κ = 0.34-0.63 vs κ = 0.17-0.20). Medians of quantitative parameters differed significantly (P < .0001 to P = .0233) between benign tissue and PCa (ADC, 1.64 × 10(-3) mm(2)/sec vs 1.13 × 10(-3) mm(2)/sec; K(trans), 0.16 min(-1) vs 0.33 min(-1); k(ep), 0.36 min(-1) vs 0.62 min(-1)). MP MR imaging has greater accuracy in the detection of recurrent PCa after RT than T2-weighted imaging alone, with no additional benefit if DCE is added to T2-weighted imaging and DW imaging.


Endothelial protein C receptor-dependent antichemotactic effects of canine protein C.
OBJECTIVE To determine whether canine protein C (CnPC) had antichemotactic effects on canine neutrophils, whether endothelial protein C receptor (EPCR) was expressed on canine neutrophils, and the role of EPCR in neutrophil chemotaxis. SAMPLE Neutrophils isolated from blood samples from healthy dogs (n = 6) and sick dogs with (2) or without (3) an inflammatory leukogram. PROCEDURES Neutrophils were analyzed by reverse transcriptase PCR assay and flow cytometry for detection of EPCR mRNA and protein expression, respectively. Neutrophils were incubated with CnPC zymogen or canine activated protein C (CnAPC), with or without RCR-379 (an anti-human EPCR antibody). Neutrophils were then allowed to migrate through a filter membrane toward a chemokine. Untreated neutrophils served as positive control samples. Migration was quantified by fluorescence measurement, and chemotaxis index (Chx) values (fluorescence of test sample/fluorescence of positive control sample) were computed. RESULTS The cDNA for EPCR was amplified, and EPCR expression was detected on neutrophil surfaces. Obtained Chx values were significantly higher in cells treated with RCR-379 than in cells treated with CnPC or CnAPC alone. The Chx values for neutrophils treated with RCR-379 were not significantly different from 1, whereas those for neutrophils treated without RCR-379 were significantly less than 1. The effects of RCR-379 on neutrophil migration were independent of concentration or activation status of protein C. CONCLUSIONS AND CLINICAL RELEVANCE Canine neutrophils expressed EPCR, and inhibition of neutrophil chemotaxis by CnPC and CnAPC depended on EPCR. Interventions with EPCR signaling may have therapeutic application in dogs.


Young offspring at genetic risk of adult psychoses: the form of the trajectory of IQ or memory may orient to the right dysfunction at the right time.
Neurocognitive dysfunctions analogous to those of adult patients have been detected in children at risk of schizophrenia and bipolar disorder. This led to the following developmental question: Do IQ and memory impairments exhibit different developmental courses from childhood to young adulthood in terms of stability or fluctuations? In a high risk sample, we used a step by step sampling approach to narrow-down the early disease mechanisms. Upstream, we started with a 20-year follow-up of 48 densely affected multigenerational kindreds, including 1500 clinically characterized adult members. We then identified 400 adult members affected by a DSM-IV schizophrenia or bipolar disorder. Downstream, we finally focused on 65 offspring (of an affected parent) aged 7 to 22, who were administered a neuropsychological battery. We then constructed cross-sectional trajectories that were compared to those of controls. The childhood IQ deficit displayed a stability until young adulthood. The delay in visual memory exhibited a non-linear two-stage trajectory: a lagging period during childhood followed by a recuperation period from adolescence until adulthood, as supported by a significant Group x Age Periods interaction. No data suggested deterioration between 7 and 22. In these offspring at genetic risk, the developmental trajectory of global IQ impairment may not apply to specific domains of cognition such as episodic memory. Different cognitive dysfunctions would mark different developmental courses. The shape of the trajectories might itself have a meaning and provide empirical leads for targeting the right dysfunction at the right time in future prevention research.


Cryptococcus neoformans resists to drastic conditions by switching to viable but non-culturable cell phenotype.
Metabolically quiescent pathogens can persist in a viable non-replicating state for months or even years. For certain infectious diseases, such as tuberculosis, cryptococcosis, histoplasmosis, latent infection is a corollary of this dormant state, which has the risk for reactivation and clinical disease. During murine cryptococcosis and macrophage uptake, stress and host immunity induce Cryptococcus neoformans heterogeneity with the generation of a sub-population of yeasts that manifests a phenotype compatible with dormancy (low stress response, latency of growth). In this subpopulation, mitochondrial transcriptional activity is regulated and this phenotype has been considered as a hallmark of quiescence in stem cells. Based on these findings, we worked to reproduce this phenotype in vitro and then standardize the experimental conditions to consistently generate this dormancy in C. neoformans. We found that incubation of stationary phase yeasts (STAT) in nutriment limited conditions and hypoxia for 8 days (8D-HYPOx) was able to produced cells that mimic the phenotype obtained in vivo. In these conditions, mortality and/or apoptosis occurred in less than 5% of the yeasts compared to 30-40% of apoptotic or dead yeasts upon incubation in normoxia (8D-NORMOx). Yeasts in 8D-HYPOx harbored a lower stress response, delayed growth and less that 1% of culturability on agar plates, suggesting that these yeasts are viable but non culturable cells (VBNC). These VBNC were able to reactivate in the presence of pantothenic acid, a vitamin that is known to be involved in quorum sensing and a precursor of acetyl-CoA. Global metabolism of 8D-HYPOx cells showed some specific requirements and was globally shut down compared to 8D-NORMOx and STAT conditions. Mitochondrial analyses showed that the mitochondrial mass increased with mitochondria mostly depolarized in 8D-HYPOx compared to 8D-NORMox, with increased expression of mitochondrial genes. Proteomic and transcriptomic analyses of 8D-HYPOx revealed that the number of secreted proteins and transcripts detected also decreased compared to 8D-NORMOx and STAT, and the proteome, secretome and transcriptome harbored specific profiles that are engaged as soon as four days of incubation. Importantly, acetyl-CoA and the fatty acid pathway involving mitochondria are required for the generation and viability maintenance of VBNC. Altogether, these data show that we were able to generate for the first time VBNC phenotype in C. neoformans. This VBNC state is associated with a specific metabolism that should be further studied to understand dormancy/quiescence in this yeast.


Coherent charge-phonon correlations and exciton dynamics in orthorhombic CH3NH3PbI3 measured by ultrafast multi-THz spectroscopy.
We use time-resolved multi-terahertz spectroscopy for the range 4-40 meV to probe coherent and incoherent ultrafast charge carrier and exciton dynamics in the low temperature orthorhombic phase of the hybrid metal halide perovskite CH3NH3PbI3. Time- and energy-resolved terahertz reflectivity maps reveal strongly damped but coherent oscillations in the 2-4 THz reststrahlen band, indicating charge coupling to a distribution of low energy phonon modes centered at 0.9 THz (3.7 meV or 30 cm-1). First-principles calculations reveal that these modes are entirely of mixed organic/inorganic sublattice character, with the power spectrum of the coherent oscillations showing a high frequency cutoff just at the onset of organic cation-only vibrations. Two anomalous reflectivity signatures are observed which are not phonon related, which we assign to a free exciton at 12 meV appearing on a 0.5 ps time scale and a defect bound exciton at 29-32 meV appearing on slower 1 ps time scale. Our measurements reveal the coherent coupling of charges to low energy vibrations of mixed sublattice character and the presence of two distinct populations of free and bound excitons at low temperatures.


Beyond Playing 20 Questions with Nature: Integrative Experiment Design in the Social and Behavioral Sciences.
The dominant paradigm of experiments in the social and behavioral sciences views an experiment as a test of a theory, where the theory is assumed to generalize beyond the experiment's specific conditions. According to this view, which Alan Newell once characterized as "playing twenty questions with nature," theory is advanced one experiment at a time, and the integration of disparate findings is assumed to happen via the scientific publishing process. In this article, we argue that the process of integration is at best inefficient, and at worst it does not, in fact, occur. We further show that the challenge of integration cannot be adequately addressed by recently proposed reforms that focus on the reliability and replicability of individual findings, nor simply by conducting more or larger experiments. Rather, the problem arises from the imprecise nature of social and behavioral theories and, consequently, a lack of commensurability across experiments conducted under different conditions. Therefore, researchers must fundamentally rethink how they design experiments and how the experiments relate to theory. We specifically describe an alternative framework, integrative experiment design, which intrinsically promotes commensurability and continuous integration of knowledge. In this paradigm, researchers explicitly map the design space of possible experiments associated with a given research question, embracing many potentially relevant theories rather than focusing on just one. The researchers then iteratively generate theories and test them with experiments explicitly sampled from the design space, allowing results to be integrated across experiments. Given recent methodological and technological developments, we conclude that this approach is feasible and would generate more-reliable, more-cumulative empirical and theoretical knowledge than the current paradigm-and with far greater efficiency.


miR-198 inhibits HIV-1 gene expression and replication in monocytes and its mechanism of action appears to involve repression of cyclin T1.
Cyclin T1 is a regulatory subunit of a general RNA polymerase II elongation factor known as P-TEFb. Cyclin T1 is also required for Tat transactivation of HIV-1 LTR-directed gene expression. Translation of Cyclin T1 mRNA has been shown to be repressed in human monocytes, and this repression is relieved when cells differentiate to macrophages. We identified miR-198 as a microRNA (miRNA) that is strongly down-regulated when monocytes are induced to differentiate. Ectopic expression of miR-198 in tissue culture cells reduced Cyclin T1 protein expression, and plasmid reporter assays verified miR-198 target sequences in the 3' untranslated region (3'UTR) of Cyclin T1 mRNA. Cyclin T1 protein levels increased when an inhibitor of miR-198 was transfected into primary monocytes, and overexpression of miR-198 in primary monocytes repressed the normal up-regulation of Cyclin T1 during differentiation. Expression of an HIV-1 proviral plasmid and HIV-1 replication were repressed in a monocytic cell line upon overexpression of miR-198. Our data indicate that miR-198 functions to restrict HIV-1 replication in monocytes, and its mechanism of action appears to involve repression of Cyclin T1 expression.


Treatment of non-AIDS progressive multifocal leukoencephalopathy with cytosine arabinoside.
This open label study determined the outcome of non-AIDS progressive multifocal leukoencephalopathy patients treated with a standard dose of intravenous cytosine arabinoside. Nineteen patients with PML proven by brain biopsy or spinal fluid polymerase chain reaction were treated with intravenous cytosine arabinoside 2 mg/kg per day for 5 days and followed for neurologic outcome by neurologic examination and MRI scanning. Seven of 19 PML patients treated with cytosine arabinoside intravenously improved neurologically. The range of follow-up for these patients was 2.0 to 4.5 years. All were left with neurologic deficits but were functionally improved, and 6 of 7 were able to independently carry out the activities of daily living. Twelve PML patients showed no evidence of response and died rapidly of their disease after treatment (range, 8 days to 6 months). All who survived their neurologic disease recovered from treatment-induced pancytopenia. Cytosine arabinoside given intravenously to non-AIDS PML patients in this small study was associated with a 36% chance of developing stabilization at 1 year. Treatment was associated with significant bone marrow toxicity. The improvement in MRI scan changes in those patients who responded took 6 weeks or longer.


Different yet similar: evolution of imprinting in flowering plants and mammals.
Genomic imprinting refers to a form of epigenetic gene regulation whereby alleles are differentially expressed in a parent-of-origin-dependent manner. Imprinting evolved independently in flowering plants and in therian mammals in association with the elaboration of viviparity and a placental habit. Despite the striking differences in plant and animal reproduction, genomic imprinting shares multiple characteristics between them. In both groups, imprinted expression is controlled, at least in part, by DNA methylation and chromatin modifications in cis-regulatory regions, and many maternally and paternally expressed genes display complementary dosage-dependent effects during embryogenesis. This suggests that genomic imprinting evolved in response to similar selective pressures in flowering plants and mammals. Nevertheless, there are important differences between plant and animal imprinting. In particular, genomic imprinting has been shown to be more flexible and evolutionarily labile in plants. In mammals, imprinted genes are organized mainly in highly conserved clusters, whereas in plants they occur in isolation throughout the genome and are affected by local gene duplications. There is a large degree of intra- and inter-specific variation in imprinted gene expression in plants. These differences likely reflect the distinct life cycles and the different evolutionary dynamics that shape plant and animal genomes.


The influence of donor-related factors on the frequency of acute cellular rejection by the recipient in the first year following heart transplantation.
This study was an attempt to identify donor-related factors influencing the frequency of acute cellular rejection episodes in cardiac transplant recipients in the first year following transplantation. The study encompassed 76 patients (14 women and 62 men) aged 6-68 years, mean 47.8+/-16.3, who received a cardiac transplant at the Institute of Cardiology in Warsaw between 2001-2005. The period of observation was from day 7 after transplantation (the day of the first biopsy by protocol) to the end of the first year. Follow-up at the outpatient department included physical examination, echocardiography, blood biochemistry and determination of circulating levels of immunosuppressive drugs. Endomyocardial biopsies were obtained and evaluated according to ISHLT criteria. The study was conducted according to the following biopsy schedule: every 7 days in the first month, every 14 days in the second, and every three months from month three to the end of the first year. The frequency of acute cellular rejection was assessed along with potential donor-related factors (age, gender, donor cause of death). The study protocol was approved by the Local Bioethics Committee of the Institute of Cardiology. Comparative statistical analysis was conducted retrospectively using the Mann-Whitney and Kruskal-Wallis tests. A total of 39 abnormal biopsies were found (degree 2 and 3 ISHLT) among the 750 biopsies performed (5.2%). No significant differences were found in the number of acute cellular rejection episodes in relation to treatment protocol and blood cyclosporine concentration in the studied period. Acute cellular rejection was occured significantly more frequently when intracerebral hemorrhage was the donor's cause of death (p<0.0131) and showed a tendency to occur more frequently when the donor was female (p<0.0611). The effects of donor age, blood type and duration of ischemia were not significant. Significantly more episodes of acute cellular rejection occur in recipients of hearts from donors who died from intracerebral hemorrhage. A trend towards more frequent episodes of acute cellular rejection appears when the donor was female.


Applications of the reflective practice questionnaire in medical education.
We sought to determine whether the Reflective Practice Questionnaire (RPQ) is a reliable measure of reflective capacity and related characteristics in medical students. We also planned to learn how the RPQ could be used in medical education. The RPQ is a 40 item self-report questionnaire that includes a multi-faceted approach to measuring reflective capacity. It also includes sub-scales on several other theoretically relevant constructs such as desire for improvement, confidence, stress, and job satisfaction. The reliabilities of reflective capacity and other sub-scales were determined by calculating their Cronbach alpha reliability values. In the present study, the RPQ was answered by 98 graduating fourth-year medical students from an American University, and these RPQ scores were compared with general public and mental health practitioner samples from a prior study using ANOVA and Bonferroni adjusted comparisons. Medical students reported a higher reflective capacity than the general public sample, but students were statistically indistinguishable from the mental health practitioner sample. For medical students, reflective capacity was associated with features of confidence, stress, and desire for improvement. Job satisfaction was positively associated with confidence in communication with patients, and negatively associated with stress when interacting with patients. A cluster analysis revealed that around 19% of the medical students exhibited a relatively high level of anxiety interacting with patients, 23% were less engaged, 5% were dissatisfied, and 7% expressed a level of over-confidence in their knowledge and skills that was concerning. The RPQ is a reliable measure of reflective capacity (Chronbach's alpha value = 0.84) and related characteristics (Cronbach's alpha values from 0.75 to 0.83) in medical students. The RPQ can be used as part of pre-post evaluations of medical education initiatives, to complement student self-reflection activities in the curriculum, and to identify students who might benefit from targeted intervention.


Fencing bodes a rapid collapse of the unique Greater Mara ecosystem.
With land privatization and fencing of thousands of hectares of communal grazing areas, East Africa is struggling with one of the most radical cultural and environmental changes in its history. The 668,500-hectare Greater Mara is of crucial importance for the great migrations of large mammals and for Maasai pastoralist culture. However, the magnitude and pace of these fencing processes in this area are almost completely unknown. We provide new evidence that fencing is appropriating land in this area at an unprecedented and accelerating speed and scale. By means of a mapped series of multispectral satellite imagery (1985-2016), we found that in the conservancies with the most fences, areal cover of fenced areas has increased with >20% since 2010. This has resulted in a situation where fencing is rapidly increasing across the Greater Mara, threatening to lead to the collapse of the entire ecosystem in the near future. Our results suggest that fencing is currently instantiating itself as a new permanent self-reinforcing process and is about to reach a critical point after which it is likely to amplify at an even quicker pace, incompatible with the region's role in the great wildebeest migration, wildlife generally, as well as traditional Maasai pastoralism.


An efficient method for gene silencing in human primary plasmacytoid dendritic cells: silencing of the TLR7/IRF-7 pathway as a proof of concept.
Plasmacytoid dendritic cells (pDC) are specialized immune cells that produce massive levels of type I interferon in response to pathogens. Unfortunately, pDC are fragile and extremely rare, rendering their functional study a tough challenge. However, because of their central role in numerous pathologies, there is a considerable need for an efficient and reproducible protocol for gene silencing in these cells. In this report, we tested six different methods for siRNA delivery into primary human pDC including viral-based, lipid-based, electroporation, and poly-ethylenimine (PEI) technologies. We show that lipid-based reagent DOTAP was extremely efficient for siRNA delivery into pDC, and did not induce cell death or pDC activation. We successfully silenced Toll-Like Receptor 7 (TLR7), CXCR4 and IFN regulatory factor 7 (IRF-7) gene expression in pDC as assessed by RT-qPCR or cytometry. Finally, we showed that TLR7 or IRF-7 silencing in pDC specifically suppressed IFN-α production upon stimulation, providing a functional validation of our transfection protocol.


Kidney stones: a fetal origins hypothesis.
Kidney stones are common, with a multifactorial etiology involving dietary, environmental, and genetic factors. In addition, patients with nephrolithiasis are at greater risk of hypertension, diabetes mellitus, metabolic syndrome, and osteoporosis, although the basis for this is not fully understood. All of these renal stone-associated conditions have also been linked with adverse early-life events, including low-birth weight, and it has been suggested that this developmental effect is due to excess exposure to maternal glucocorticoids in utero. This is proposed to result in long-term increased hypothalamic-pituitary-axis activation; there are mechanisms through which this effect could also promote urinary lithogenic potential. We therefore hypothesize that the association between renal stone disease and hypertension, diabetes mellitus, metabolic syndrome, and osteoporosis may be related by a common pathway of programming in early life, which, if validated, would implicate the developmental origins hypothesis in the etiology of nephrolithiasis.


Treatment of olecranon bursitis in horses: 10 cases (1986-1993).
Medical records of 10 horses with olecranon bursitis were reviewed to examine treatments, evaluate a technique for en bloc resection of the bursa in standing horses, and determine outcome of the horses after treatment. Before admission, 6 horses had been treated by needle aspiration of fluid from the mass, followed by injection of corticosteroids. Subsequent treatment for 2 of these 6 horses included open drainage and packing of the cavity with gauze soaked in 7% iodine solution. None resolved after these treatments. After admission to the hospital, 5 horses were treated medically and 5 were treated by en bloc resection of the bursa. One horse that had received intralesional injection of a radionuclide was lost to follow-up evaluation. One horse treated conservatively by open drainage and packing and 1 treated by injection of a radionuclide had resolution of the olecranon bursitis. Only 1 of these 2 horses had a cosmetic result. The acquired bursae decreased in size in 2 horses (1 treated with a corticosteroid and 1 with orgotein), but were still visible 7 and 46 months after treatment, respectively. The surgery site of 4 horses that were treated by en bloc resection healed by primary intention, and the owners of these horses were pleased with the cosmetic results. The suture line of 1 horse dehisced 5 days after surgery. Proliferative granulation tissue was removed on 2 occasions, and the site healed by second intention after 2 months. A small knot and some white hair remained at the surgery site.


Socioeconomic differences in lack of recreational walking among older adults: the role of neighbourhood and individual factors.
People with a low socioeconomic status (SES) are more likely to be physically inactive than their higher status counterparts, however, the mechanisms underlying this socioeconomic gradient in physical inactivity remain largely unknown. Our aims were (1) to investigate socioeconomic differences in recreational walking among older adults and (2) to examine to what extent neighbourhood perceptions and individual cognitions regarding regular physical activity can explain these differences. Data were obtained by a large-scale postal survey among a stratified sample of older adults (age 55-75 years) (N = 1994), residing in 147 neighbourhoods of Eindhoven and surrounding areas, in the Netherlands. Multilevel logistic regression analyses assessed associations between SES (i.e. education and income), perceptions of the social and physical neighbourhood environment, measures of individual cognitions derived from the Theory of Planned Behaviour (e.g. attitude, perceived behaviour control), and recreational walking for >/=10 minutes/week (no vs. yes). Participants in the lowest educational group (OR 1.67 (95% CI, 1.18-2.35)) and lowest income group (OR 1.40 (95% CI, 0.98-2.01)) were more likely to report no recreational walking than their higher status counterparts. The association between SES and recreational walking attenuated when neighbourhood aesthetics was included in the model, and largely reduced when individual cognitions were added to the model (with largest effects of attitude, and intention regarding regular physical activity). The assiation between poor neighbourhood aesthetics and no recreational walking attenuated to (borderline) insignificance when individual cognitions were taken into account. Both neighbourhood aesthetics and individual cognitions regarding physical activity contributed to the explanation of socioeconomic differences in no recreational walking. Neighbourhood aesthetics may explain the association between SES and recreational walking largely via individual cognitions towards physical activity. Intervention and policy strategies to reduce socioeconomic differences in lack of recreational walking among older adults would be most effective if they intervene on both neighbourhood perceptions as well as individual cognitions.


RNA Interference based Approach to Down Regulate Osmoregulators of Whitefly (Bemisia tabaci): Potential Technology for the Control of Whitefly.
Over the past decade RNA interference (RNAi) technology has emerged as a successful tool not only for functional genomics, but in planta expression of short interfering RNAs (siRNAs) that could offer great potential for insect pest management. The diet of insects feeding exclusively on phloem sieves contains water and sugars as main components, and the uptake of the liquid food greatly depends on the osmotic pressure within the insect body. Based on this physiological mechanism, transgenic plants of Nicotiana tabacum were generated expressing double stranded RNA (dsRNA) against both aquaporin (AQP) and a sucrase gene, alpha glucosidase (AGLU). These two genes are involved in osmotic pressure maintenance particularly in sap sucking insects, and the aim was to disrupt osmoregulation within the insect ultimately leading to mortality. Real time quantitative PCR (RT-qPCR) was performed to assess the suppression of gene expression in Bemisia tabaci (B. tabaci) and mortality was recorded during transgenic tobacco feeding bioassays. Feeding of insects on plants expressing dsRNA significantly reduced the transcript level of the target genes in B. tabaci after six days of feeding and more than 70% mortality was observed in B. tabaci fed on transgenic plants compared to the control plants. Our data shows that down-regulation of genes related to osmoregulation may find practical applications for the control of this important pest in cotton and other crops.


Risk equalisation in voluntary health insurance markets: A three country comparison.
The paper summarises the conclusions for health policy from the experience of three countries who have introduced risk equalisation subsidies, in their voluntary health insurance (VHI) markets. The countries chosen are Australia, Ireland and South Africa. All of these countries have developed VHI markets and have progressed towards introducing risk equalisation. The objective of such subsidies is primarily to make VHI affordable while encouraging efficiency in health care production. The paper presents a conceptual framework to understand and compare risk equalisation subsidies in VHI markets. The paper outlines how such subsidies are organised in each of the countries and identifies problems that arise in their implementation. We conclude that the objectives of risk equalisation, in VHI markets are no different to those in countries with mandatory insurance systems. We find that the introduction of risk equalisation subsidies is complex and that countries seeking to introduce risk equalisation in VHI markets must carefully consider how such subsidies advance their overall health policy goals. Furthermore, we conclude that such subsidies must be structured correctly as otherwise incentives exist for risk selection which may threaten affordability and efficiency. Our overall conclusion is that also in voluntary health insurance markets risk equalisation has a role in meeting the related public policy objectives of risk solidarity and affordability, and without it these objectives are severely undermined.


Microscopic analysis of MTT stained boar sperm cells.
The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI) Stations is limited.


Interfacing Membrane Mimetics with Mass Spectrometry.
Membrane proteins play critical physiological roles and make up the majority of drug targets. Due to their generally low expression levels and amphipathic nature, membrane proteins represent challenging molecular entities for biophysical study. Mass spectrometry offers several sensitive approaches to study the biophysics of membrane proteins. By preserving noncovalent interactions in the gas phase and using collisional activation to remove solubilization agents inside the mass spectrometer, native mass spectrometry (MS) is capable of studying isolated assemblies that would be insoluble in aqueous solution, such as membrane protein oligomers and protein-lipid complexes. Conventional methods use detergent to solubilize the protein prior to electrospray ionization. Gas-phase activation inside the mass spectrometer removes the detergent to yield the isolated proteins with bound ligands. This approach has proven highly successful for ionizing membrane proteins. With the appropriate choice of detergents, membrane proteins with bound lipid species can be observed, which allows characterization of protein-lipid interactions. However, detergents have several limitations. They do not necessarily replicate the native lipid bilayer environment, and only a small number of protein-lipid interactions can be resolved. In this Account, we summarize the development of different membrane mimetics as cassettes for MS analysis of membrane proteins. Examples include amphipols, bicelles, and picodiscs with a special emphasis on lipoprotein nanodiscs. Polydispersity and heterogeneity of the membrane mimetic cassette is a critical issue for study by MS. Ever more complex data sets consisting of overlapping protein charge states and multiple lipid-bound entities have required development of new computational, theoretical, and experimental approaches to interpret both mass and ion mobility spectra. We will present the rationale and limitations of these approaches. Starting with the early work on empty nanodiscs, we chart developments that culminate in recent high-resolution studies of membrane protein-lipid complexes ejected from nanodiscs. For the latter, increasing collision energies allowed progressive removal of nanodisc components, beginning with the scaffold proteins and continuing through successive shells of lipids, allowing direct characterization of the stoichiometry of the annular lipid belt that surrounds the membrane protein. We consider future directions for the study of membrane proteins in membrane mimetics, including the development of mixed lipid systems and native bilayer environments. Unambiguous assignment of these heterogeneous systems will rely heavily upon further enhancements in both data analysis protocols and instrumental resolution. We anticipate that these developments will provide new insights into the factors that control dynamic protein-lipid interactions in a variety of tailored and natural lipid environments.


Insight into the molecular mechanisms of protein stabilizing osmolytes from global force-field variations.
A prominent class of osmolytes that are able to stabilize proteins in their native fold consist of small highly water-soluble molecules with a large dipole moment and hydrophobic groups attached to the positively charged end of the molecule, for which we coin the term dipolar/hydrophobic osmolytes. For TMAO, which is a prime member of this class, we perform large-scale water-explicit MD simulations and determine the bulk solution activity coefficient as well as the affinity to a stretched polyglycine chain for varying TMAO dipolar strength and hydrophobicity. Double optimization with respect to experimental values for the activity coefficient and the polyglycine transfer free energy is achieved. The resulting optimal TMAO force field shows excellent transferability to different concentrations and also reproduces transfer free energies of various amino acids, including the tryptophan anomaly, for which TMAO acts as a denaturant. By globally analyzing the thermodynamic and structural properties of suboptimal TMAO force fields, we identify the frustration between dipolar and hydrophobic interactions as the working mechanism and the design principle of dipolar/hydrophobic osmolytes.


Pre-hospital intubation by anaesthesiologists in patients with severe trauma: an audit of a Norwegian helicopter emergency medical service.
Anaesthesiologists are airway management experts, which is one of the reasons why they serve as pre-hospital emergency physicians in many countries. However, limited data are available on the actual quality and safety of anaesthesiologist-managed pre-hospital endotracheal intubation (ETI). To explore whether the general indications for ETI are followed and what complications are recorded, we analysed the use of pre-hospital ETI in severely traumatised patients treated by anaesthesiologists in a Norwegian helicopter emergency medical service (HEMS). A retrospective audit of prospectively registered data concerning patients with trauma as the primary diagnosis and a National Committee on Aeronautics score of 4 - 7 during the period of 1994-2005 from a mixed rural/urban Norwegian HEMS was performed. Among the 1255 cases identified, 238 successful pre-hospital ETIs out of 240 attempts were recorded (99.2% success rate). Furthermore, we identified 47 patients for whom ETI was performed immediately upon arrival to the emergency department (ED). This group represented 16% of all intubated patients. Of the ETIs performed in the ED, 43 patients had an initial Glasgow Coma Score (GCS) < 9. Compared to patients who underwent ETI in the ED, patients who underwent pre-hospital ETI had significantly lower median GCS (3 (3-6) vs. 6 (4-8)), lower revised trauma scores (RTS) (3.8 (1.8-5.9) vs. 5.0 (4.1-6.0)), longer mean scene times (23 +/- 13 vs. 11 +/- 11 min) and longer mean transport times (22 +/- 16 vs. 13 +/- 14 min). The audit also revealed that very few airway management complications had been recorded. We found a very high success rate of pre-hospital ETI and few recorded complications in the studied anaesthesiologist-manned HEMS. However, a substantial number of trauma patients were intubated first on arrival in the ED. This delay may represent a quality problem. Therefore, we believe that more studies are needed to clarify the reasons for and possible clinical consequences of the delayed ETIs.


Worldwide frequency distribution of the 'Gait keeper' mutation in the DMRT3 gene.
For centuries, domestic horses have represented an important means of transport and served as working and companion animals. Although their role in transportation is less important today, many horse breeds are still subject to intense selection based on their pattern of locomotion. A striking example of such a selected trait is the ability of a horse to perform additional gaits other than the common walk, trot and gallop. Those could be four-beat ambling gaits, which are particularly smooth and comfortable for the rider, or pace, used mainly in racing. Gaited horse breeds occur around the globe, suggesting that gaitedness is an old trait, selected for in many breeds. A recent study discovered that a nonsense mutation in DMRT3 has a major impact on gaitedness in horses and is present at a high frequency in gaited breeds and in horses bred for harness racing. Here, we report a study of the worldwide distribution of this mutation. We genotyped 4396 horses representing 141 horse breeds for the DMRT3 stop mutation. More than half (2749) of these horses also were genotyped for a SNP situated 32 kb upstream of the DMRT3 nonsense mutation because these two SNPs are in very strong linkage disequilibrium. We show that the DMRT3 mutation is present in 68 of the 141 genotyped horse breeds at a frequency ranging from 1% to 100%. We also show that the mutation is not limited to a geographical area, but is found worldwide. The breeds with a high frequency of the stop mutation (>50%) are either classified as gaited or bred for harness racing.


The more often you see an object, the easier it becomes to track it.
Is it easier to track objects that you have seen repeatedly? We compared repeated blocks, where identities were the same from trial to trial, to unrepeated blocks, where identities varied. People were better in tracking objects that they saw repeatedly. We tested four hypotheses to explain this repetition benefit. First, perhaps the repeated condition benefits from consistent mapping of identities to target and distractor roles. However, the repetition benefit persisted even when both the repeated and the unrepeated conditions used consistent mapping. Second, repetition might improve the ability to recover targets that have been lost, or swapped with distractors. However, we observed a larger repetition benefit for color-color conjunctions, which do not benefit from such error recovery processes, than for unique features, which do. Furthermore, a repetition benefit was observed even in the absence of distractors. Third, perhaps repetition frees up resources by reducing memory load. However, increasing memory load by masking identities during the motion phase reduced the repetition benefit. The fourth hypothesis is that repetition facilitates identity tracking, which in turn improves location tracking. This hypothesis is consistent with all our results. Thus, our data suggest that identity and location tracking share a common resource.


The utility of the Hopkins Verbal Learning Test (Chinese version) for screening dementia and mild cognitive impairment in a Chinese population.
The Hopkins Verbal Learning Test (HVLT) has been validated for detecting dementia in English-speaking populations. However, no studies have examined the Chinese version of the HVLT scale, and appropriate cut-off scores for dementia in the Chinese population remain unclear. 631 subjects aged 60 and over were recruited at a memory clinic at Dongzhimen Hospital in Beijing. Of these, 249 were classified as exhibiting normal cognition (NC), 134 were diagnosed with mild cognitive impairment (MCI), 97 were diagnosed with Alzheimer's disease (AD), 14 met the diagnosis for vascular dementia (VaD), and 50 were diagnosed with other types of dementia, including mixed dementia. The discriminative capacity of the HVLT total learning score, recognition score and total score were calculated to determine their sensitivity and specificity for detecting MCI, AD and other dementias, and various cut-off scores. HVLT scores were affected by age, education and sex. The HVLT total learning score exhibited an optimal balance between sensitivity and specificity using a cut-off score of 15.5 for distinguishing AD and other types of dementia from NC using the ROC curve, with sensitivity of 94.7% for distinguishing AD and all types of dementia, and specificity of 92.5% for detecting AD and 93.4% for detecting all types of dementias. We stratified the AD and MCI groups by age, and calculated the validity in each age group. In the 50-64 years age group, when the cutoff score was 18.5, the sensitivity of 0.955 and specificity of 0.921 were obtained for discriminating the NC and AD groups, and in the 65-80 years group, and optimal sensitivity and specificity values (0.948 and 0.925, respectively) were obtained with a cutoff score of 14.5. When the cutoff score was 21.5 in HVLT total recall, an optimal balance was obtained between sensitivity and specificity (69.1% and 70.7%, respectively) in distinguishing MCI from NC. A cut-off score of 15.5 in the HVLT total learning score led to high discriminative capacity between the dementia and NC groups. This suggests that the HVLT total learning score can provide a useful tool for discriminating dementia, but not MCI, from NC in clinical and epidemiological practice.


Possible association between SNAP-25 single nucleotide polymorphisms and alterations of categorical fluency and functional MRI parameters in Alzheimer's disease.
Synaptosomal-associated protein of 25 kDa (SNAP-25) is an age-regulated vesicular SNARE protein involved in the exocytosis of neurotransmitters from synapses, a process that is altered in Alzheimer's disease (AD). Changes in SNAP-25 levels are suggested to contribute to age-related decline of cognitive function, and single nucleotide polymorphisms (SNPs) in the SNAP-25 gene are present in neuropsychiatric conditions and play a role in determining IQ phenotypes. To verify a possible role of SNAP-25 in AD, we analyzed five gene polymorphisms in patients with AD (n = 607), replicating the study in subjects with amnestic mild cognitive impairment (aMCI) (n = 148) and in two groups of age-matched healthy controls (HC1: n = 615 and HC2: n = 310). Results showed that the intronic rs363050 (A) and rs363043 (T) alleles, as well as the rs363050/rs363043 A-T haplotype are significantly more frequent in AD and aMCI and are associated with pathological scores of categorical fluency in AD. Notably, functional MRI analyses indicated that SNAP-25 genotypes correlate with a significantly decreased brain activity in the cingulate cortex and in the frontal (middle and superior gyri) and the temporo-parietal (angular gyrus) area. SNAP-25 polymorphisms may be associated with AD and correlate with alterations in categorical fluency and a reduced localized brain activity. SNAP-25 polymorphisms could be used as surrogate markers for the diagnosis of AD and of cognitive deficit; these SNPs might also have a possible predictive role in the natural history of AD.


Charge compensation in trivalent cation doped bulk rutile TiO2.
Doping of TiO(2) is a very active field, with a particularly large effort expended using density functional theory (DFT) to model doped TiO(2); this interest has arisen from the potential for doping to be used in tuning the band gap of TiO(2) for photocatalytic applications. Doping is also of importance for modifying the reactivity of an oxide. Finally, dopants can also be unintentionally incorporated into an oxide during processing, giving unexpected electronic properties. To unravel properly how doping impacts on the properties of a metal oxide requires a modelling approach that can describe such systems consistently. Unfortunately, DFT, as used in the majority of studies, is not suitable for application here and in many cases cannot even yield a qualitatively consistent description. In this paper we investigate the doping of bulk rutile TiO(2) with trivalent cations, Al, Ga and In, using DFT, DFT corrected for on-site Coulomb interactions (DFT + U, with U on oxygen 2p states) and hybrid DFT (the screened exchange HSE06 exchange correlation functional) in an effort to better understand the performance of DFT in describing such fundamental doping scenarios and to analyse the process of charge compensation with these dopants. With all dopants, DFT delocalizes the oxygen hole polaron that results from substitution of Ti with the lower valence cation. DFT also finds an undistorted geometry and does not produce the characteristic polaron state in the band gap. DFT + U and hybrid DFT both localize the polaron, and this is accompanied by a distortion to the structure around the oxygen hole site. DFT + U and HSE06 both give a polaron state in the band gap. The band gap underestimation present in DFT + U means that the offset of the gap state from both the valence and the conduction band cannot be properly described, while the hybrid DFT offsets should be correct. We have investigated dopant charge compensation by formation of oxygen vacancies. Due to the large number of calculations required, we use DFT + U for these studies. We find that the most stable oxygen vacancy site has either a very small positive formation energy or is negative, so under typical experimental conditions, anion vacancy formation will compensate for the dopant.


Hepatoprotective herbal drug, silymarin from experimental pharmacology to clinical medicine.
Silymarin, a flavonolignan from 'milk thistle' (Silybum marianum) plant is used almost exclusively for hepatoprotection and amounts to 180 million US dollars business in Germany alone. In this review we discuss about its safety, efficacy and future uses in liver diseases. The use of silymarin may replace the polyherbal formulations and will avoid the major problems of standardization, quality control and contamination with heavy metals or bacterial toxins. Silymarin consists of four flavonolignan isomers namely--silybin, isosilybin, silydianin and silychristin. Among them, silybin being the most active and commonly used. Silymarin is orally absorbed and is excreted mainly through bile as sulphates and conjugates. Silymarin offers good protection in various toxic models of experimental liver diseases in laboratory animals. It acts by antioxidative, anti-lipid peroxidative, antifibrotic, anti-inflammatory, membrane stabilizing, immunomodulatory and liver regenerating mechanisms. Silymarin has clinical applications in alcoholic liver diseases, liver cirrhosis, Amanita mushroom poisoning, viral hepatitis, toxic and drug induced liver diseases and in diabetic patients. Though silymarin does not have antiviral properties against hepatitis virus, it promotes protein synthesis, helps in regenerating liver tissue, controls inflammation, enhances glucuronidation and protects against glutathione depletion. Silymarin may prove to be a useful drug for hepatoprotection in hepatobiliary diseases and in hepatotoxicity due to drugs. The non traditional use of silymarin may make a breakthrough as a new approach to protect other organs in addition to liver. As it is having a good safety profile, better patient tolerability and an effective drug at an affordable price, in near future new derivatives or new combinations of this drug may prove to be useful.


Retinal gene delivery by adeno-associated virus (AAV) vectors: Strategies and applications.
Adeno-associated virus (AAV) vectors are the most widely used vehicle systems for neuronal gene transfer. This popularity is based on the non-pathogenic nature of AAVs and their versatility making them a multifunctional vector system for basic research and clinical applications. AAVs are successfully applied in clinical and pre-clinical gene therapy studies for inherited retinal disorders. Their excellent transduction profile and efficiency also boosted the use of AAV vectors in basic research. The AAV vector system can be easily modified and adjusted at multiple levels to allow for optimized and specific gene expression in target cells. Here, we will provide an overview on the AAV vector system and its applications focusing on gene transfer into retinal cells. Furthermore, we will outline and discuss strategies for the optimization of AAV gene transfer by modifications to the AAV vector expression cassette, the AAV capsid or the routes of vector administration.


Comparative genomics of recent Shiga toxin-producing Escherichia coli O104:H4: short-term evolution of an emerging pathogen.
The large outbreak of diarrhea and hemolytic uremic syndrome (HUS) caused by Shiga toxin-producing Escherichia coli O104:H4 in Europe from May to July 2011 highlighted the potential of a rarely identified E. coli serogroup to cause severe disease. Prior to the outbreak, there were very few reports of disease caused by this pathogen and thus little known of its diversity and evolution. The identification of cases of HUS caused by E. coli O104:H4 in France and Turkey after the outbreak and with no clear epidemiological links raises questions about whether these sporadic cases are derived from the outbreak. Here, we report genome sequences of five independent isolates from these cases and results of a comparative analysis with historical and 2011 outbreak isolates. These analyses revealed that the five isolates are not derived from the outbreak strain; however, they are more closely related to the outbreak strain and each other than to isolates identified prior to the 2011 outbreak. Over the short time scale represented by these closely related organisms, the majority of genome variation is found within their mobile genetic elements: none of the nine O104:H4 isolates compared here contain the same set of plasmids, and their prophages and genomic islands also differ. Moreover, the presence of closely related HUS-associated E. coli O104:H4 isolates supports the contention that fully virulent O104:H4 isolates are widespread and emphasizes the possibility of future food-borne E. coli O104:H4 outbreaks. In the summer of 2011, a large outbreak of bloody diarrhea with a high rate of severe complications took place in Europe, caused by a previously rarely seen Escherichia coli strain of serogroup O104:H4. Identification of subsequent infections caused by E. coli O104:H4 raised questions about whether these new cases represented ongoing transmission of the outbreak strain. In this study, we sequenced the genomes of isolates from five recent cases and compared them with historical isolates. The analyses reveal that, in the very short term, evolution of the bacterial genome takes place in parts of the genome that are exchanged among bacteria, and these regions contain genes involved in adaptation to local environments. We show that these recent isolates are not derived from the outbreak strain but are very closely related and share many of the same disease-causing genes, emphasizing the concern that these bacteria may cause future severe outbreaks.


Comparative effects of osmotic and secretory diarrhoea on brush-border disaccharide hydrolases in rat.
The effects of diarrhoea on the activities of brush-border disaccharidases namely lactase (EC 126.96.36.199), maltase (EC 220.127.116.11) and sucrase (EC 184.108.40.206) of Sprague-Dawley strain albino rats were induced in the rats with mannitol while secretory diarrhoea was induced with Salmonella typhimurium after an initial treatment with streptomycin. The activities of the enzymes were significantly reduced by diarrhoea. The extent of reduction in enzyme activity varied in the different segment of the small intestine in all the groups. The jejuno-ileal region had more changes in enzyme activities than in the duodenum. Higher activity levels were observed for maltase than for lactase. In the osmotic diarrhoea model, lactase activity was significantly lowered (P < 0.05) in the experimental group from day 5 to 10. Maltase activity on the other hand was significantly lowered (P < 0.001) at the peak of diarrhoea. Sucrase activity was also lowered significantly (P < 0.025) in the experimental animals within the first 10 days of diarrhoeal induction. In the secretory diarrhoea model, lactase activity was similar in all the experimental groups except for the streptomycin-salmonella-treated groups and control (P < 0.05). Higher lactase activity levels were observed in the secretory diarrhoea model compared to level in the osmotic diarrhoea model. Maltase activity levels were also lowered significantly (P < 0.05) in the experimental animals. Streptomycin had no effect on the activity of maltase.


Depressed Langerhans cell migration and reduced contact hypersensitivity response in mice lacking TNF receptor p75.
Epidermal Langerhans cells (LC) belong to the dendritic cell family and represent the major APC within the skin. LC capture epicutaneous Ag, migrate into regional lymph nodes, and present Ag to T cells, thereby initiating primary immune response. The migratory properties of LC are an essential component of their function. The molecular mechanisms responsible for LC migration are far less defined. However, evidence has been accumulating to suggest that TNF-alpha, a major proinflammatory cytokine, plays an important role in promoting DC migration. To confirm the role of TNF-alpha in LC migration and to examine which type of TNF receptor signaling is involved in such an event, we utilized gene-targeted knockout mice lacking TNF receptor p55 or p75. The migration of LC was assessed by examining the frequency of hapten-bearing cells in draining lymph nodes following hapten FITC painting, and the accumulation of dendritic cells in draining lymph nodes after intradermal injection of TNF-alpha. While LC migration was normal in p55-deficient mice, the migration was markedly depressed in p75-deficient mice. Receptor p75-deficient mice also demonstrated a hyporesponsiveness in allergen-induced contact dermatitis, but a normal responsiveness in irritant-induced contact dermatitis. These results suggest that p75-dependent signaling plays a crucial role in the migration of LC and in the initiation of cutaneous immune responses.


Multimorbidity patterns and association with mortality in 0.5 million Chinese adults.
Few studies have assessed the relationship between multimorbidity patterns and mortality risk in the Chinese population. We aimed to identify multimorbidity patterns and examined the associations of multimorbidity patterns and the number of chronic diseases with the risk of mortality among Chinese middle-aged and older adults. We used data from the China Kadoorie Biobank and included 512,723 participants aged 30 to 79 years. Multimorbidity was defined as the presence of two or more of the 15 chronic diseases collected by self-report or physical examination at baseline. Multimorbidity patterns were identified using hierarchical cluster analysis. Cox regression was used to estimate the associations of multimorbidity patterns and the number of chronic diseases with all-cause and cause-specific mortality. Overall, 15.8% of participants had multimorbidity. The prevalence of multimorbidity increased with age and was higher in urban than rural participants. Four multimorbidity patterns were identified, including cardiometabolic multimorbidity (diabetes, coronary heart disease, stroke, and hypertension), respiratory multimorbidity (tuberculosis, asthma, and chronic obstructive pulmonary disease), gastrointestinal and hepatorenal multimorbidity (gallstone disease, chronic kidney disease, cirrhosis, peptic ulcer, and cancer), and mental and arthritis multimorbidity (neurasthenia, psychiatric disorder, and rheumatoid arthritis). During a median of 10.8 years of follow-up, 49,371 deaths occurred. Compared with participants without multimorbidity, cardiometabolic multimorbidity (hazard ratios [HR] = 2.20, 95% confidence intervals [CI]: 2.14 - 2.26) and respiratory multimorbidity (HR = 2.13, 95% CI:1.97 - 2.31) demonstrated relatively higher risks of mortality, followed by gastrointestinal and hepatorenal multimorbidity (HR = 1.33, 95% CI:1.22 - 1.46). The mortality risk increased by 36% (HR = 1.36, 95% CI: 1.35 - 1.37) with every additional disease. Cardiometabolic multimorbidity and respiratory multimorbidity posed the highest threat on mortality risk and deserved particular attention in Chinese adults.


An upstream open reading frame is essential for feedback regulation of ascorbate biosynthesis in Arabidopsis.
Ascorbate (vitamin C) is an essential antioxidant and enzyme cofactor in both plants and animals. Ascorbate concentration is tightly regulated in plants, partly to respond to stress. Here, we demonstrate that ascorbate concentrations are determined via the posttranscriptional repression of GDP-l-galactose phosphorylase (GGP), a major control enzyme in the ascorbate biosynthesis pathway. This regulation requires a cis-acting upstream open reading frame (uORF) that represses the translation of the downstream GGP open reading frame under high ascorbate concentration. Disruption of this uORF stops the ascorbate feedback regulation of translation and results in increased ascorbate concentrations in leaves. The uORF is predicted to initiate at a noncanonical codon (ACG rather than AUG) and encode a 60- to 65-residue peptide. Analysis of ribosome protection data from Arabidopsis thaliana showed colocation of high levels of ribosomes with both the uORF and the main coding sequence of GGP. Together, our data indicate that the noncanonical uORF is translated and encodes a peptide that functions in the ascorbate inhibition of translation. This posttranslational regulation of ascorbate is likely an ancient mechanism of control as the uORF is conserved in GGP genes from mosses to angiosperms.


Six Values Never to Silence: Jewish Perspectives on Nazi Medical Professionalism.
An ideological case study based on medical profession norms during the Third Reich will be used to exemplify the importance of diversity in the manifestations of professional ethics. The German professional medical community banned their Jewish colleagues from treating German citizens. This included legally mandated employment discrimination and outright censure which led to a professional ethic devoid of diverse voices. While the escalation to the T-4 program and medicalized genocide was influenced by many causes, the intentional, ethnocentric-based exclusion of voices was an important contributing element to the chronicled degradation of societal mores. For illustration, six core Jewish values-life, peace, justice, mercy, scholarship, and sincerity of intention-will be detailed for their potential to inspire health-care professionals to defend and protect minorities and for readers to think critically about the role of medical professionalism in Third Reich society. The Jewish teachings highlight the inherent professional obligations physicians have toward their patients in contrast to the Third Reich's corruption of patient-centered professionalism. More fundamentally, juxtaposing Jewish and Nazi teachings exposes the loss of perspective when a profession's identity spurns diversity. To ensure respect for persons in all vulnerable minorities, the first step is addressing professional inclusion of minority voices.


A prolactin-dependent sexually dimorphic mechanism of migraine chronification.
Determination of possible sex differences in mechanisms promoting migraine progression and the contribution of prolactin and the prolactin long (PRLR-L) and short (PRLR-S) receptor isoforms. The majority of patients with chronic migraine and medication overuse headache are female. Prolactin is present at higher levels in women and increases migraine. Prolactin signaling at the PRLR-S selectively sensitizes nociceptors in female rodents, while expression of the PRLR-L is protective. Medication overuse headache was modeled by repeated sumatriptan administration in male and female mice. Periorbital and hindpaw cutaneous allodynia served as a surrogate of migraine-like pain. PRLR-L and PRLR-S isoforms were measured in the trigeminal ganglion with western blotting. Possible co-localization of PRLR with serotonin 5HT1B and 5HT1D receptors was determined with RNAscope. Cabergoline, a dopamine receptor agonist that inhibits circulating prolactin, was co-administered with sumatriptan. Nasal administration of CRISPR/Cas9 plasmid was used to edit expression of both PRLR isoforms. PRLR was co-localized with 5HT1B or 5HT1D receptors in the ophthalmic region of female trigeminal ganglion. A single injection of sumatriptan increased serum PRL levels in female mice. Repeated sumatriptan promoted cutaneous allodynia in both sexes but down-regulated trigeminal ganglion PRLR-L, without altering PRLR-S, only in females. Co-administration of sumatriptan with cabergoline prevented allodynia and down-regulation of PRLR-L only in females. CRISPR/Cas9 editing of both PRLR isoforms in the trigeminal ganglion prevented sumatriptan-induced periorbital allodynia in females. We identified a sexually dimorphic mechanism of migraine chronification that involves down-regulation of PRLR-L and increased signaling of circulating prolactin at PRLR-S. These studies reveal a previously unrecognized neuroendocrine mechanism linking the hypothalamus to nociceptor sensitization that increases the risk of migraine pain in females and suggest opportunities for novel sex-specific therapies including gene editing through nasal delivery of CRISPR/Cas9 constructs.


Transcorneal Electrical Stimulation Reduces Neurodegenerative Process in a Mouse Model of Glaucoma.
Glaucoma is a neurodegenerative disease in which the retinal ganglion cell axons of the optic nerve degenerate concomitant with synaptic changes in the retina, leading finally to death of the retinal ganglion cells (RGCs). Electrical stimulation has been used to improve neural regeneration in a variety of systems, including in diseases of the retina. Therefore, the focus of this study was to investigate whether transcorneal electrical stimulation (TES) in the DBA2/J mouse model of glaucoma could improve retinal or optic nerve pathology and serve as a minimally invasive treatment option. Mice (10 months-old) received 21 sessions of TES over 8 weeks, after which we evaluated RGC number, axon number, and anterograde axonal transport using histology and immunohistochemistry. To gain insight into the mechanism of proposed protection, we also evaluated inflammation by quantifying CD3+ T-cells and Iba1+ microglia; perturbations in metabolism were shown via the ratio pAMPK to AMPK, and changes in trophic support were tested using protein capillary electrophoresis. We found that TES resulted in RGC axon protection, a reduction in inflammatory cells and their activation, improved energy homeostasis, and a reduction of the cell death-associated p75NTR. Collectively, the data indicated that TES maintained axons, decreased inflammation, and increased trophic factor support, in the form of receptor presence and energy homeostasis, suggesting that electrical stimulation impacts several facets of the neurodegenerative process in glaucoma.


African American women's experiences with the initial discovery, diagnosis, and treatment of breast cancer.
To describe the experiences of African American women living with breast cancer following the primary diagnosis and while undergoing initial treatment. Phenomenologic. 13 African American women (ages 30-66) purposefully selected from two oncology clinics in the mid-South. Phenomenologic interviews (transcribed verbatim) and field notes were analyzed using Colaizzi's method of phenomenologic description and analysis. Experience Trajectory, Femininity, and Spirituality were the three major themes. The Experience Trajectory subthemes were finding the lump, getting the diagnosis, undergoing surgery and adjuvant treatment. The Femininity subthemes were loss of all or part of the breast, loss of hair, and sexual attractiveness to a man. Spirituality was reflected as a reliance on God. Telling the story of their experience trajectory during their breast cancer experience is valuable in assessing African American women's feelings, emotions, and fears of body changes that occur during surgery and treatment. Their spirituality helps them through this experience. Research involving both African American women and their partners would provide greater insight into specific relationship patterns and communication related to sexuality during this experience. Nurses need to listen to the stories of African American women about the initial experience of discovery, diagnosis, and treatment of breast cancer so they can be more informed advocates for these women. African American women need more information from healthcare providers regarding the whole experience trajectory.


Secretory Carcinoma of the Lacrimal Gland: A Rare Case Report.
Secretory carcinoma is a salivary gland malignancy that recapitulates secretory carcinoma of the breast, along with its shared ETV6-NTRK3 gene fusion. Characterization of histopathologic, immunohistochemical, and molecular genetic features of this neoplasm has led to reclassification of a heterogeneous group of salivary gland carcinomas as secretory carcinoma and to identification of this neoplasm in other gland-containing tissues. The authors describe a 52-year-old man who presented with a 2-week history of diplopia and a well-circumscribed right orbital mass. The tumor was resected via lateral orbitotomy approach. Pathologic evaluation demonstrated secretory carcinoma, previously not described in the main lacrimal gland. Recognition of lacrimal gland secretory carcinoma may lead to reappraisal of morphologically similar, but biologically heterogeneous lacrimal gland neoplasms, providing an insight into this tumor's clinical presentation and prognosis. Accurate diagnosis of this malignancy has important management and prognostic implications, particularly with emergence of targeted therapies.


Changing how we risk-categorise in Ramadan: Does the IDF-DAR scoring system achieve the requirements for people with diabetes internationally?
The IDF-DAR guidance was most recently updated this year. The most notable change is the moving to a scoring system from a tabulated risk categorization to determine and classify the risk of harm from fasting derived from an online survey. This change may be appealing and is welcomed. However, such a system and the methodology underpinning it is not without limitations. This commentary highlights some of these limitations and the associated limited safe options available to individuals with diabetes desiring to fast during Ramadan. Overlooked clinical considerations that deserve formal recognition include the role of technology (aspects relating to glucose monitoring and/or insulin delivery) and previous experience of safe Ramadan fasting. Further, duration of fast (which can almost double in temperate regions from winter to summer) needs greater emphasis. We also advocate separate scoring systems for people with type 1 diabetes and complex type 2 diabetes. The guidance acknowledges fasting is an individual's decision, however the general message needs to be more person-centred and currently only presents a binary approach to fasting - all or nothing choices. We propose and discuss addition options including, trial fasting of voluntary fasts, starting the fast and terminating due to health and/or safety, intermittent fasting and winter fasting.


In vitro characterization of T-type calcium channel antagonist TTA-A2 and in vivo effects on arousal in mice.
T-type calcium channels have been implicated in many behaviorally important neurophysiological processes, and altered channel activity has been linked to the pathophysiology of neurological disorders such as insomnia, epilepsy, Parkinson's disease, depression, schizophrenia, and pain. We have previously identified a number of potent and selective T-type channel antagonists (Barrow et al., 2007; Shipe et al., 2008; Yang et al., 2008). Here we describe the properties of the antagonist TTA-A2 [2-(4-cyclopropylphenyl)-N-((1R)-1-{5-[(2,2,2-trifluoroethyl)oxo]-pyridin-2-yl}ethyl)acetamide], assessed in patch-clamp experiments. TTA-A2 blocks T-type channels (Ca(v)3.1, 3.2, 3.3) voltage dependently and with high potency (IC(50) ∼100 nM). Stimulation at 3 Hz revealed additional use dependence of inhibition. A hyperpolarized shift of the channel availability curve and delayed channel recovery from inactivation suggest that the compound preferentially interacts with and stabilizes inactivated channels. The compound showed a ∼300-fold selectivity for Ca(v)3 channels over high-voltage activated calcium channels. Inhibitory effects on native T-type currents were confirmed in brain slice recordings from the dorsal lateral geniculate nucleus and the subthalamic nucleus. Furthermore, we demonstrate that in vivo T-type channel inhibition by TTA-A2 suppresses active wake and promotes slow-wave sleep in wild-type mice but not in mice lacking both Ca(v)3.1 and Ca(v)3.3, suggesting the selective effect of TTA-A2 on recurrent thalamocortical network activity. The discovery of the potent and selective T-type channel antagonist TTA-A2 has enabled us to study the in vivo effects of pharmacological T-channel inhibition on arousal in mice, and it will help to explore the validity of these channels as potential drug targets for sleep-related and other neurological diseases.


Comparison of the sociodemographic characteristics of the large NutriNet-Santé e-cohort with French Census data: the issue of volunteer bias revisited.
A recurring concern in traditional and in Web-based studies pertains to non-representativeness due to volunteer bias. We investigated this issue in an ongoing, large population-based e-cohort. The sample included 122 912 individuals enrolled in the Internet-based, nutrition-focused NutriNet-Santé study between May 2009 and March 2014, with complete baseline data. Participants were recruited via recurrent multimedia campaigns and other traditional and online strategies. Individuals aged 18+ years, residing in France and having Internet access, were eligible for enrolment. Their sociodemographic characteristics were compared with the corresponding 2009 Census data via χ(2) goodness-of-fit tests. The effectiveness of statistical weighting of the e-cohort data was also explored. The sample exhibited marked geographical and sociodemographic diversity, including volunteers belonging to typically under-represented subgroups in traditional surveys (unemployed, immigrants, the elderly). Nonetheless, the proportions of women, relatively well-educated individuals and those who are married or cohabiting, were notably larger compared with the corresponding national figures (women: 78.0% vs 52.4%; postsecondary education: 61.5% vs 24.9%; married or cohabiting: 70.8% vs 62.0%, respectively; all p<0.0001). There were notable sociodemographic differences between the general French population and this general population-based e-cohort, some of which were corrected by statistical weighting. The findings bear on the potential generalisability of future investigations in the context of e-epidemiology.